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Identification of pathways regulating cell size and cell-cycle progression by RNAi.

机译：通过RNAi鉴定调节细胞大小和细胞周期进程的途径。

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摘要

Many high-throughput loss-of-function analyses of the eukaryotic cell cycle have relied on the unicellular yeast species Saccharomyces cerevisiae and Schizosaccharomyces pombe. In multicellular organisms, however, additional control mechanisms regulate the cell cycle to specify the size of the organism and its constituent organs. To identify such genes, here we analysed the effect of the loss of function of 70% of Drosophila genes (including 90% of genes conserved in human) on cell-cycle progression of S2 cells using flow cytometry. To address redundancy, we also targeted genes involved in protein phosphorylation simultaneously with their homologues. We identify genes that control cell size, cytokinesis, cell death and/or apoptosis, and the G1 and G2/M phases of the cell cycle. Classification of the genes into pathways by unsupervised hierarchical clustering on the basis of these phenotypes shows that, in addition to classical regulatory mechanisms such as Myc/Max, Cyclin/Cdk and E2F, cell-cycle progression in S2 cells is controlled by vesicular and nuclear transport proteins, COP9 signalosome activity and four extracellular-signal-regulated pathways (Wnt, p38betaMAPK, FRAP/TOR and JAK/STAT). In addition, by simultaneously analysing several phenotypes, we identify a translational regulator, eIF-3p66, that specifically affects the Cyclin/Cdk pathway activity.
机译：真核细胞周期的许多高通量功能丧失分析都依赖于单细胞酵母菌酿酒酵母和粟酒裂殖酵母。然而，在多细胞生物中，附加的控制机制调节细胞周期以指定生物及其组成器官的大小。为了鉴定此类基因，我们在这里使用流式细胞仪分析了70％的果蝇基因（包括90％的人类保守基因）功能丧失对S2细胞的细胞周期进程的影响。为了解决冗余问题，我们还针对与蛋白质磷酸化有关的基因及其同源物。我们确定控制细胞大小，细胞分裂，细胞死亡和/或凋亡，以及细胞周期的G1和G2 / M期的基因。根据这些表型，通过无监督的层次聚类将基因分类为途径，结果表明，除了经典的调控机制（例如Myc / Max，Cyclin / Cdk和E2F）之外，S2细胞中的细胞周期进程还受到水泡和核仁的控制。转运蛋白，COP9信号体活性和四种细胞外信号调节途径（Wnt，p38betaMAPK，FRAP / TOR和JAK / STAT）。此外，通过同时分析几种表型，我们确定了一个翻译调节因子eIF-3p66，它特异性地影响Cyclin / Cdk通路的活性。

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